High molecular weight DNA assembly in vivo for synthetic biology applications

Abstract

DNA assembly is the key technology of the emerging interdisciplinary field of synthetic biology. While the assembly of smaller DNA fragments is usually performed in vitro, high molecular weight DNA molecules are assembled in vivo via homologous recombination in the host cell. Escherichia coli, Bacillus subtilis and Saccharomyces cerevisiae are the main hosts used for DNA assembly in vivo. Progress in DNA assembly over the last few years has paved the way for the construction of whole genomes. This review provides an update on recent synthetic biology advances with particular emphasis on high molecular weight DNA assembly in vivo in E. coli, B. subtilis and S. cerevisiae. Special attention is paid to the assembly of whole genomes, such as those of the first synthetic cell, synthetic yeast and minimal genomes.

• Bacillus subtilis
• Escherichia coli
• genome assembly
• homologous recombination
• minimal genome
• Saccharomyces cerevisiae
• synthetic cell
• synthetic yeast

Acknowledgements

Cordial thanks to all researchers who contributed to these advances in DNA assembly. M.J. and J.W.A. are supported by the Engineering and Physical Sciences Research Council (EPSRC).

Declaration of interest

There are no conflicts of interest associated with this manuscript.